An inducible model for unraveling the effects of advanced glycation end-product accumulation in aging connective tissues

ABSTRACT

Purpose

In connective tissues there is a clear link between increasing age and degeneration. Advanced glycation end-products (AGEs) are believed to play a central role. AGEs are sugar-induced non-enzymatic crosslinks which accumulate in collagen with age and diabetes, altering tissue mechanics and cellular function. Despite ample correlative evidence linking collagen glycation to tissue degeneration, little is known how AGEs impact cell-matrix interactions, limiting therapeutic options. One reason for this limited understanding is that AGEs are typically induced using high concentrations of ribose which decrease cell viability, making it impossible to investigate cell-matrix interactions. The objective of this study was to develop a system to trigger AGE accumulation while maintaining cell viability.

Materials and Methods

Using cell-seeded high density collagen gels, we investigated the effect of two systems for AGE induction, ribose at low concentrations (30, 100, and 200 mM) over 15 days of culture and riboflavin (0.25 and 0.75 mM) induced with blue light for 40 seconds (riboflavin-465 nm).

Results

We found ribose and riboflavin-465 nm treatment produces fluorescent AGE quantities which match and/or exceed human fluorescent AGE levels for various tissues, ages, and diseases, without affecting cell viability or metabolism. Interestingly, a 40 second treatment of riboflavin-465 nm produced similar levels of fluorescent AGEs as 3 days of 100 mM ribose treatment.

Conclusions

Riboflavin-465 nm is a promising method to trigger AGEs on demand in vivo or in vitro without impacting cell viability and offers potential for unraveling the mechanism of AGEs in age and diabetes related tissue damage.

KEYWORDS:

• Advanced glycation end-products (AGEs)
• aging
• crosslinking
• ribose
• riboflavin
• collagen

Acknowledgments

The authors would like to thank are Dr. Rebecca Heise, Dr. Michael Valentine, Dr. Carl Sayer, Dr. Barbara Boyan, and Cydney Dennis for their support in this research. The authors acknowledge the use of facilities within the Nanomaterials Characterization Core at Virginia Commonwealth University.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Supplementary material

Supplemental data for this article can be accessed here

Additional information

Funding

This study was supported by PI start-up funds and this research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.